Safety evaluation on defatted soybean particles after nanofabrication

Main Article Content

Shenguang Tong
Hongjiang Hao
Xiaofei Li
Hongbo Li
Zhiyong Liao
Zhihua Wu
Hongbing Chen


nano defatted soybean particles, safety evaluation, acute toxicity, histopathological examination, gastrointestinal distribution, cytotoxicity assay


Many nanoparticles are used in food for increasing the bioavailability of nutrients. Nano defatted soybean particles (nDSPs) were promising as nanoparticles of a traditional food, but its safety remains pending. In this work, the possible toxicity of nDSP was tested on cell and mouse models. Cell proliferation and the viability of defatted soybean particles (DSPs), DSP tracking in gastrointestinal, and tissue histopathological examination were performed. The Zeta potential of nDSP was as low as ?16 ± 3 mV and had no cytotoxicity on Caco-2 cells or animal models. In the gastrointestinal tract, the nDSP showed similar absorption patterns with DSP of 500 nm or 1 ?m. In acute toxicity assessment, no abnormal behavior was observed in mice after DSP administration, and no noticeable tissue damage and inflammatory lesion were found either. Here, we show that DSPs, including nDSP, are safe at a single dose of 10 g/kg body weight, regardless of the particle size. The food property and aggregation behavior both help to make the nanoparticle safe.

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Abudayyak M, Gurkaynak TA, Ozhan G. In vitro evaluation of cobalt oxide nanoparticle-induced toxicity. Toxicol Indust Health. 2017;33:646–54. 10.1177/0748233717706633
Bihari P, Vippola M, Schultes S, Praetner M, Khandoga AG, Reichel CA, et al. Optimized dispersion of nanoparticles for biological in vitro and in vivo studies. Part Fibre Toxicol. 2008;5:14. http://
Bitencourt PER, Ferreira LM, Cargnelutti LO, Denardi L, Boligon A, Fleck M, et al. A new biodegradable polymeric nanoparticle formulation containing Syzygium cumini: Phytochemical profile, antioxidant and antifungal activity and in vivo toxicity. Indust Crops Products. 2016;83:400–7. indcrop.2016.01.007
Boyles MS, Ranninger C, Reischl R, Rurik M, Tessadri R, Kohlbacher O, et al. Copper oxide nanoparticle toxicity profiling using untargeted metabolomics. Part Fibre Toxicol. 2016;13:49.
Brenner SA, Neu-Baker NM, Caglayan C, Zurbenko IG. Occupational exposure to airborne nanomaterials: An assessment of worker exposure to aerosolized metal oxide nanoparticles in semiconductor wastewater treatment. J Occup Environ Hyg. 2015;12:469–81.
Cai K, Hou Y, Hu Y, Zhao L, Luo Z, Shi Y, et al. Correlation of the cytotoxicity of TiO2 nanoparticles with different particle sizes on a sub-200-nm scale. Small. 2011;7:3026–31. http://dx.doi. org/10.1002/smll.201101170
Cui Y, Liu H, Zhou M, Duan Y, Li N, Gong X, et al. Signaling path-way of inflammatory responses in the mouse liver caused by TiO2 nanoparticles. J Biomed Mater Res A. 2011;96:221–9.
Frey A, Giannasca KT, Weltzin R, Giannasca PJ, Reggio H, Lencer WI, et al. Role of the glycocalyx in regulating access of microparticles to apical plasma membranes of intestinal epithelial cells: Implications for microbial attachment and oral vaccine targeting. J Exp Med. 1996;184:1045–59. http://dx.doi. org/10.1084/jem.184.3.1045
Gandamalla D, Lingabathula H, Yellu N. Nano titanium exposure induces dose- and size-dependent cytotoxicity on human epithelial lung and colon cells. Drug Chem Toxicol. 2019;42(1):24– 34.
Graham UM, Jacobs G, Yokel RA, Davis BH, Dozier AK, Birch ME, et al. From dose to response: In vivo nanoparticle processing and potential toxicity. Adv Exp Med Biol. 2017;947:71–100. http://
Hillyer JF, Albrecht RM. Gastrointestinal persorption and tissue distribution of differently sized colloidal gold nanoparticles. J Pharm Sci. 2001;90:1927–36. jps.1143
Hinkley GK, Carpinone P, Munson JW, Powers KW, Roberts  SM. Oral absorption of PEG-coated versus uncoated gold nanospheres: Does agglomeration matter? Part Fibre Toxicol. 2015;12:9.
Jani PU, Florence AT, Mccarthy DE. Further histological evidence of the gastrointestinal absorption of polystyrene nanospheres in the rat. Int J Pharmaceut. 1992;84:245–52. http://dx.doi. org/10.1016/0378-5173(92)90162-U
Jeevanandam J, Barhoum A, Chan YS, Dufresne A, Danquah MK. Review on nanoparticles and nanostructured materials: History, sources, toxicity and regulations. Beilstein J Nanotechnol. 2018;9:1050–74.
Jia XL, Li SY, Dang SS, Cheng YA, Zhang X, Wang WJ, et al. Increased expression of chondroitin sulphate proteoglycans in rat hepatocellular carcinoma tissues. World J Gastroenterol. 2012;18:3962–76.
Lacroix IME, Chen XM, Kitts DD, Li-Chan ECY. Investigation into the bioavailability of milk protein-derived peptides with dipeptidylpeptidase IV inhibitory activity using Caco-2 cell monolayers. Food Funct. 2017;8:701–9.
Lanone S, Boczkowski J. Biomedical applications and potential health risks of nanomaterials: Molecular mechanisms. Curr Mol Med. 2006;6:651–63.
Leung YH, Yung MM, Ng AM, Ma AP, Wong SW, Chan CM, et al. Toxicity of CeO2 nanoparticles – The effect of nanoparticle
properties. J Photochem Photobiol B. 2015;145:48–59. http://
L’Hocine L, Boye JI. Allergenicity of soybean: New developments in identification of allergenic proteins, cross-reactivities and hypoallergenization technologies. Crit Rev Food Sci Nutr. 2007;47:127–43.
Li JQ, Zou XW, Li CL, Zhong JH, Chen Y, Zhang XY, et al. Expression of novel cancer/testis antigen TMEM31 increases during metastatic melanoma progression. Oncol Lett. 2017;13:2269–73.
Liao CD, Hung WL, Lu WC, Jan KC, Shih DYC, Yeh AI, et al. Differential tissue distribution of sesaminol triglucoside and its metabolites in rats fed with lignan glycosides from sesame meal with or without nano/submicrosizing. J Agric Food Chem. 2010;58:563–9.
Lopes VR, Loitto V, Audinot JN, Bayat N, Gutleb AC, Cristobal S. Dose-dependent autophagic effect of titanium dioxide nanoparticles in human HaCaT cells at non-cytotoxic J Nanobiotechnol. 2016;14:22. s12951-016-0174-0
Lusas EW, Riaz MN. Soy protein products: Processing and use. J Nutr. 1995;125:573S–80S.
Ma JQ, Guan RF, Shen HT, Lu F, Xiao CG, Liu MQ, et al. Comparison of anticancer activity between lactoferrin nanoliposome and lacto-ferrin in Caco-2 cells in vitro. Food Chem Toxicol. 2013;59:72–7.
Maurer LL, Yang XY, Schindler AJ, Taggart RK, Jiang CJ, Hsu-Kim H, et  al. Intracellular trafficking pathways in silver nanoparticle uptake and toxicity in Caenorhabditis elegans. Nanotoxicology. 2016;10:831–835.
Meggs WJ, Brewer KL. Weight gain associated with chronic exposure to chlorpyrifos in rats. J Med Toxicol. 2007;3:89–93. http://
Michael B, Yano B, Sellers RS, Perry R, Morton D, Roome N, et al. Evaluation of organ weights for rodent and non-rodent toxicity studies: A review of regulatory guidelines and a survey of current practices. Toxicol Pathol. 2007;35:742–50. http://dx.doi. org/10.1080/01926230701595292
Nohynek GJ, Dufour EK, Roberts MS. Nanotechnology, cosmetics and the skin: Is there a health risk? Skin Pharmacol Physiol. 2008;21:136–49.
Pakrashi S, Tan C, Wang WX. Bioaccumulation-based silver nanoparticle toxicity in Daphnia magna and maternal impacts. Environ Toxicol Chem. 2017;36:3359–66. http://dx.doi. org/10.1002/etc.3917
Parhi R, Suresh P. Preparation and characterization of solid lipid nanoparticles – A review. Curr Drug Discov Technol. 2012;9:2– 16.
Park EJ, Lee GH, Yoon C, Jeong U, Kim Y, Chang J, et al. Tissue distribution following 28 day repeated oral administration of aluminum-based nanoparticles with different properties and the in vitro toxicity. J Appl Toxicol. 2017;37:1408–19. http://dx.doi. org/10.1002/jat.3509
Patel MR, San Martin-Gonzalez MF. Characterization of ergocalciferol loaded solid lipid nanoparticles. J Food Sci. 2012;77:N8–13.
Qu DF, Gu YP, Feng LF, Han JZ. High content analysis technology for evaluating the joint toxicity of sunset yellow and sodium sulfite in vitro. Food Chem. 2017;233:135–43. http://dx.doi. org/10.1016/j.foodchem.2017.04.102
Recordati C, De Maglie M, Bianchessi S, Argentiere S, Cella C, Mattiello S, et al. Tissue distribution and acute toxicity of silver after single intravenous administration in mice: Nanospecific and size-dependent effects. Part Fibre Toxicol. 2016;13:12.
Singh RP, Ramarao P. Cellular uptake, intracellular trafficking and cytotoxicity of silver nanoparticles. Toxicol Lett. 2012;213:249– 59.
Smith JN, Thomas DG, Jolley H, Kodali VK, Littke MH, Munusamy P, et al. All that is silver is not toxic: Silver ion and particle kinetics reveals the role of silver ion aging and dosimetry on the toxicity of silver nanoparticles. Part Fibre Toxicol. 2018;15:47. http://dx.
Tang QJ, Tao KZ, Yun L, Sun XJ, Geng MY, Jiang CL. Immunocytochemical localization of secretory component in Paneth cell secretory granules-rat Paneth cells participate in acquired immunity. J Mol Histol. 2005;36:331–5. http://dx.doi. org/10.1007/s10735-005-9003-8
Teo S, Stirling D, Thomas S, Hoberman A, Kiorpes  A, Khetani  V. A 90-day oral gavage toxicity study of D-methylphenidate and D,L-methylphenidate in Sprague-Dawley rats. Toxicology. 2002;179:183– 96.
Ugwah-Oguejiofor CJ, Okoli CO, Ugwah MO, Umaru ML, Ogbulie CS, Mshelia HE, et al. Acute and subacute toxicity of aqueous extract of aerial parts of Caralluma dalzielii N. E. Brown in mice and rats. Heliyon. 2019;5:e01179. heliyon.2019.e01179
van Pomeren M, Peijnenburg W, Brun NR, Vijver MG. A novel experimental and modelling strategy for nanoparticle toxicity testing enabling the use of small quantities. Int J Environ Res Public Health. 2017;14:1348. ijerph14111348
Vance ME, Kuiken T, Vejerano EP, McGinnis SP, Hochella MF Jr., Rejeski D, et al. Nanotechnology in the real world: Redeveloping the nanomaterial consumer products inventory. Beilstein J Nanotechnol. 2015;6:1769–80.
Wang T, Qin GX, Sun ZW, Zhao Y. Advances of research on glycinin and beta-conglycinin: A review of two major soybean allergenic proteins. Crit Rev Food Sci Nutr. 2014;54:850–62. http://
Wang X, Ji Z, Chang CH, Zhang H, Wang M, Liao YP, et al. Use of coated silver nanoparticles to understand the relationship of particle dissolution and bioavailability to cell and lung toxicological potential. Small. 2014;10:385–98. smll.201301597
Wassef L, Quadro L. Uptake of dietary retinoids at the maternal-fetal barrier: In vivo evidence for the role of lipoprotein lipase and alternative pathways. J Biol Chem. 2011;286:32198–207.
Wu Z, Chen H, Wu L, Yang A, Li X. The method for nano defatted soybean particles manufacture. SIPO of China, ZL 2009 1 0132261.8, 2011.
Wu Z, Lin Q, Mao H, Yang A, Chen H. The particle size and aggregation of soybean powder after milling. Food Sci (Chinese). 2008;29:315–17.
Zhang Y, Chen X, Zhao B, Wu H, Yuan L, Zhang H, et al. Biosafety study and mechanism comparison on two types of silica with different nanostructures. Toxicol Res (Camb). 2017;6:487–98.
Zhao J, Castranova V. Toxicology of nanomaterials used in nanomedicine. J Toxicol Environ Health B Crit Rev. 2011;14:593–632.
Zhao Y, Wang C, Chow AH, Ren K, Gong T, Zhang Z, et al. Self-nanoemulsifying drug delivery system (SNEDDS) for oral delivery of Zedoary essential oil: Formulation and bioavailability studies. Int J Pharm. 2010;383:170–7.